Antioxidant enzymes suppress nitric oxide production through the inhibition of NF-κB activation: Role of H2O2 and nitric oxide in inducible nitric oxide synthase expression in macrophages

Yoon Ju Han; Young Guen Kwon; Hun Taeg Chung; Sung Ki Lee; Richard L. Simmons; Timothy R. Billiar; Young Myeong Kim

doi:10.1006/niox.2001.0367

Antioxidant enzymes suppress nitric oxide production through the inhibition of NF-κB activation: Role of H₂O₂ and nitric oxide in inducible nitric oxide synthase expression in macrophages

Yoon Ju Han, Young Guen Kwon, Hun Taeg Chung, Sung Ki Lee, Richard L. Simmons, Timothy R. Billiar, Young Myeong Kim

Research output: Contribution to journal › Article › peer-review

90 Citations (Scopus)

Abstract

Reactive molecules O₂^-, H₂O₂, and nitrogen monoxide (NO) are produced from macrophages following exposure to lipopolysaccharide (LPS) and involved in cellular signaling for gene expression. Experiments were carried out to determine whether these molecules regulate inducible nitric oxide synthase (iNOS) gene expression in RAW264.7 macrophages exposed to LPS. NO production was inhibited by the antioxidative enzymes catalase, horseradish peroxidase, and myeloperoxidase but not by superoxide dismutase (SOD). In contrast, the NO-producing activity of LPS-stimulated RAW264.7 cells was enhanced by the NO scavengers hemoglobin (Hb) and myoglobin. The antioxidant enzymes decreased levels of iNOS mRNA and protein in LPS-stimulated RAW264.7 cells, whereas the NOS inhibitor N^G-monomethyl-L-argine as well as Hb increased the level of iNOS protein but not mRNA, indicating that NO inhibits iNOS protein expression. NF-κB was activated in LPS-stimulated RAW264.7 cells and the activation was significantly inhibited by antioxidant enzymes, but not by Hb. Similar results were obtained using LPS-stimulated rodent peritoneal macrophages. Extracellular O₂^- generation by LPS-stimulated macrophages was suppressed by SOD, but not by antioxidative enzymes, while accumulation of intracellular reactive oxygen species was inhabited by antioxidative enzymes, but not by SOD. Exogenous H₂O₂ induced NF-κB activation in macrophages, which was inhibited by catalase and pyrroline dithaocarbamate (PDTC). H₂O₂ enhanced iNOS expression and NO production in peritoneal macrophages when added with interferon-γ, and the effect of H₂O₂ was inhibited by catalase and PDTC. These findings suggest that H₂O₂ production from LPS-stimulated macrophages participates in the upregulation of iNOS expression via NF-κB activation and that NO is a negative feedback inhibitor of iNOS protein expression.

Original language	English
Pages (from-to)	504-513
Number of pages	10
Journal	Nitric Oxide - Biology and Chemistry
Volume	5
Issue number	5
DOIs	https://doi.org/10.1006/niox.2001.0367
Publication status	Published - 2001

Bibliographical note

Funding Information:
This work was supported by Vascular System Research Grant of KOSEF (Y.M.K.), Korea Research Foundation Grant 2000-2-22200-001-3 (S.K.L), Ministry of Health and Welfare Grant HMP-99-N-01-0001 (Y.G.K), and NIH Grant R01-GM-44100 (T.R.B).

All Science Journal Classification (ASJC) codes

Biochemistry
Physiology
Clinical Biochemistry
Cancer Research

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This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1006/niox.2001.0367

Cite this

Han, Y. J., Kwon, Y. G., Chung, H. T., Lee, S. K., Simmons, R. L., Billiar, T. R., & Kim, Y. M. (2001). Antioxidant enzymes suppress nitric oxide production through the inhibition of NF-κB activation: Role of H₂O₂ and nitric oxide in inducible nitric oxide synthase expression in macrophages. Nitric Oxide - Biology and Chemistry, 5(5), 504-513. https://doi.org/10.1006/niox.2001.0367

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title = "Antioxidant enzymes suppress nitric oxide production through the inhibition of NF-κB activation: Role of H2O2 and nitric oxide in inducible nitric oxide synthase expression in macrophages",

abstract = "Reactive molecules O2-, H2O2, and nitrogen monoxide (NO) are produced from macrophages following exposure to lipopolysaccharide (LPS) and involved in cellular signaling for gene expression. Experiments were carried out to determine whether these molecules regulate inducible nitric oxide synthase (iNOS) gene expression in RAW264.7 macrophages exposed to LPS. NO production was inhibited by the antioxidative enzymes catalase, horseradish peroxidase, and myeloperoxidase but not by superoxide dismutase (SOD). In contrast, the NO-producing activity of LPS-stimulated RAW264.7 cells was enhanced by the NO scavengers hemoglobin (Hb) and myoglobin. The antioxidant enzymes decreased levels of iNOS mRNA and protein in LPS-stimulated RAW264.7 cells, whereas the NOS inhibitor NG-monomethyl-L-argine as well as Hb increased the level of iNOS protein but not mRNA, indicating that NO inhibits iNOS protein expression. NF-κB was activated in LPS-stimulated RAW264.7 cells and the activation was significantly inhibited by antioxidant enzymes, but not by Hb. Similar results were obtained using LPS-stimulated rodent peritoneal macrophages. Extracellular O2- generation by LPS-stimulated macrophages was suppressed by SOD, but not by antioxidative enzymes, while accumulation of intracellular reactive oxygen species was inhabited by antioxidative enzymes, but not by SOD. Exogenous H2O2 induced NF-κB activation in macrophages, which was inhibited by catalase and pyrroline dithaocarbamate (PDTC). H2O2 enhanced iNOS expression and NO production in peritoneal macrophages when added with interferon-γ, and the effect of H2O2 was inhibited by catalase and PDTC. These findings suggest that H2O2 production from LPS-stimulated macrophages participates in the upregulation of iNOS expression via NF-κB activation and that NO is a negative feedback inhibitor of iNOS protein expression.",

author = "Han, {Yoon Ju} and Kwon, {Young Guen} and Chung, {Hun Taeg} and Lee, {Sung Ki} and Simmons, {Richard L.} and Billiar, {Timothy R.} and Kim, {Young Myeong}",

note = "Funding Information: This work was supported by Vascular System Research Grant of KOSEF (Y.M.K.), Korea Research Foundation Grant 2000-2-22200-001-3 (S.K.L), Ministry of Health and Welfare Grant HMP-99-N-01-0001 (Y.G.K), and NIH Grant R01-GM-44100 (T.R.B).",

year = "2001",

doi = "10.1006/niox.2001.0367",

language = "English",

volume = "5",

pages = "504--513",

journal = "Nitric Oxide - Biology and Chemistry",

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Han, YJ, Kwon, YG, Chung, HT, Lee, SK, Simmons, RL, Billiar, TR & Kim, YM 2001, 'Antioxidant enzymes suppress nitric oxide production through the inhibition of NF-κB activation: Role of H₂O₂ and nitric oxide in inducible nitric oxide synthase expression in macrophages', Nitric Oxide - Biology and Chemistry, vol. 5, no. 5, pp. 504-513. https://doi.org/10.1006/niox.2001.0367

Antioxidant enzymes suppress nitric oxide production through the inhibition of NF-κB activation: Role of H₂O₂ and nitric oxide in inducible nitric oxide synthase expression in macrophages. / Han, Yoon Ju; Kwon, Young Guen; Chung, Hun Taeg et al.
In: Nitric Oxide - Biology and Chemistry, Vol. 5, No. 5, 2001, p. 504-513.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Antioxidant enzymes suppress nitric oxide production through the inhibition of NF-κB activation

T2 - Role of H2O2 and nitric oxide in inducible nitric oxide synthase expression in macrophages

AU - Han, Yoon Ju

AU - Kwon, Young Guen

AU - Chung, Hun Taeg

AU - Lee, Sung Ki

AU - Simmons, Richard L.

AU - Billiar, Timothy R.

AU - Kim, Young Myeong

N1 - Funding Information: This work was supported by Vascular System Research Grant of KOSEF (Y.M.K.), Korea Research Foundation Grant 2000-2-22200-001-3 (S.K.L), Ministry of Health and Welfare Grant HMP-99-N-01-0001 (Y.G.K), and NIH Grant R01-GM-44100 (T.R.B).

PY - 2001

Y1 - 2001

N2 - Reactive molecules O2-, H2O2, and nitrogen monoxide (NO) are produced from macrophages following exposure to lipopolysaccharide (LPS) and involved in cellular signaling for gene expression. Experiments were carried out to determine whether these molecules regulate inducible nitric oxide synthase (iNOS) gene expression in RAW264.7 macrophages exposed to LPS. NO production was inhibited by the antioxidative enzymes catalase, horseradish peroxidase, and myeloperoxidase but not by superoxide dismutase (SOD). In contrast, the NO-producing activity of LPS-stimulated RAW264.7 cells was enhanced by the NO scavengers hemoglobin (Hb) and myoglobin. The antioxidant enzymes decreased levels of iNOS mRNA and protein in LPS-stimulated RAW264.7 cells, whereas the NOS inhibitor NG-monomethyl-L-argine as well as Hb increased the level of iNOS protein but not mRNA, indicating that NO inhibits iNOS protein expression. NF-κB was activated in LPS-stimulated RAW264.7 cells and the activation was significantly inhibited by antioxidant enzymes, but not by Hb. Similar results were obtained using LPS-stimulated rodent peritoneal macrophages. Extracellular O2- generation by LPS-stimulated macrophages was suppressed by SOD, but not by antioxidative enzymes, while accumulation of intracellular reactive oxygen species was inhabited by antioxidative enzymes, but not by SOD. Exogenous H2O2 induced NF-κB activation in macrophages, which was inhibited by catalase and pyrroline dithaocarbamate (PDTC). H2O2 enhanced iNOS expression and NO production in peritoneal macrophages when added with interferon-γ, and the effect of H2O2 was inhibited by catalase and PDTC. These findings suggest that H2O2 production from LPS-stimulated macrophages participates in the upregulation of iNOS expression via NF-κB activation and that NO is a negative feedback inhibitor of iNOS protein expression.

AB - Reactive molecules O2-, H2O2, and nitrogen monoxide (NO) are produced from macrophages following exposure to lipopolysaccharide (LPS) and involved in cellular signaling for gene expression. Experiments were carried out to determine whether these molecules regulate inducible nitric oxide synthase (iNOS) gene expression in RAW264.7 macrophages exposed to LPS. NO production was inhibited by the antioxidative enzymes catalase, horseradish peroxidase, and myeloperoxidase but not by superoxide dismutase (SOD). In contrast, the NO-producing activity of LPS-stimulated RAW264.7 cells was enhanced by the NO scavengers hemoglobin (Hb) and myoglobin. The antioxidant enzymes decreased levels of iNOS mRNA and protein in LPS-stimulated RAW264.7 cells, whereas the NOS inhibitor NG-monomethyl-L-argine as well as Hb increased the level of iNOS protein but not mRNA, indicating that NO inhibits iNOS protein expression. NF-κB was activated in LPS-stimulated RAW264.7 cells and the activation was significantly inhibited by antioxidant enzymes, but not by Hb. Similar results were obtained using LPS-stimulated rodent peritoneal macrophages. Extracellular O2- generation by LPS-stimulated macrophages was suppressed by SOD, but not by antioxidative enzymes, while accumulation of intracellular reactive oxygen species was inhabited by antioxidative enzymes, but not by SOD. Exogenous H2O2 induced NF-κB activation in macrophages, which was inhibited by catalase and pyrroline dithaocarbamate (PDTC). H2O2 enhanced iNOS expression and NO production in peritoneal macrophages when added with interferon-γ, and the effect of H2O2 was inhibited by catalase and PDTC. These findings suggest that H2O2 production from LPS-stimulated macrophages participates in the upregulation of iNOS expression via NF-κB activation and that NO is a negative feedback inhibitor of iNOS protein expression.

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